Semitransparent slide and filter combination for a microscope

ABSTRACT

A slide is arranged to be semitransparent to scatter the light emitted from a microscope to optically eliminate the outlines of pores of a filter. Material such as cell, nuclei or extracellular materials is captured on the top surface of the filter by filtration. A specimen preparation can be made by placing the filter with the material thereon on the semitransparent slide. A metal evaporation layer, so-called multicoating, can be positioned on the surface of a cover glass, so that the material can be more clearly recognized, especially at high magnification.

This application is a continuation of U.S. application Ser. No.07/476,484 filed as PCT/JP89/00729, Jul. 20, 1989, now abandoned.

TECHNICAL FIELD

This invention relates to a slide which is made to be semitransparent, acover glass whose top surface is multicoated, and a method of preparinga specimen preparation. By placing a filter, which, in advance, hascollected material such as cells by filtration, on a semitransparentslide, a specimen preparation can be prepared.

BACKGROUND ART

In the past, a preparation is usually made by smearing material such ascells on a slide glass. According to this method, the material iscaptured by excoriation or centesis and then smeared on a slide glass.

Then the slide glass with the material thereon is preserved in, forexample, dyeing solution. Thereafter a mounting medium is appliedthereon to seal the material, and a cover glass is placed thereon.

In this prior art, the work to smear the material on the slide glass iscompleted by hand, thus the material could easily be destroyed anddeformed. Further, when the material is preserved in the solution, muchof it drops from the slide glass into the solution. Also, a lot ofexpreience and skill are required to accomplish this smear method.

To make the preparation easier, Japanese Patent Application No.61-250188, for example, introduces a new method wherein a suspensionmedium including material is drawn through a polycarbonate membranefilter, which is transparent, has a thickness about 5 to 10 μm andinnumerable microscopic pores are created through the filter. After thematerial is captured on the filter as the suspension medium is drawnthrough the pores, a number of needed solutions such as dyeing solutionsare applied to the material.

According to this new method, since it is unnecessary to smear thematerial on the slide glass, the problem that the material is destroyedand deformed can be eased and the outlines of the material can better berecognized as they naturally are.

After the dyeing process is completed, the material is supposed to beplaced on a slide glass to prepare a specimen preparation. To make thepreparation with this new method, however, either the material has to betransferred to the slide glass (see U.S. Pat. No. 4,395,493), or thefilter with the material thereon should be placed on the slide glass andthe filter has to be melted by a melting liquid, eliminating the poresand leaving the material on the slide glass.

When the material collected on the filter is transferred on the slideglass, which is done by pressing the filter onto the slide glass as totranscribe the material to the slide glass, much of it is destroyed anddeformed by the pressure. Further, by this so-called transcribingprocess, it is possible that only a small percentage of the wholematerial can be transcribed onto the slide glass. As far as the othermethod is concerned, the material would be shrunken and faded by themelting liquid.

Concerning these problems, it is desirable that a specimen preparationcan be prepared only by placing a filter with material thereon onto aslide glass, eliminating the need for transcribing and meltingprocesses. However, when a specimen preparation is prepared by thismethod, the outlines of the pores of the filter are recognized, and theydouble with the outlines of the material, so that an examination of thematerial will be inaccurate.

Therefore, the major object of the present invention is to opticallyeliminate the pores of the filter, to eliminating the need for thetranscribing and melting processes, to provide a slide (not necessarilya glass slide) and a cover glass that enable you to accurately examinethe material, and to provide a method of preparing a specimenpreparation using the semitransparent slide.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a perspective view of a preferred embodiment of asemitransparent slide according to the present invention.

FIG. 2 is a perspective view of another preferred embodiment of asemitransparent slide according to the present invention.

FIG. 3 is a sectional view of another preferred embodiment of asemitransparent slide according to the present invention.

FIG. 4 is a sectional view of a preparation made by a method ofpreparing a specimen preparation according to the present invention.

FIG. 5 is a sectional view of a preferred embodiment of a cover glassaccording to the present invention.

DISCLOSURE OF INVENTION

To accomplish the object of the present invention, a slide 1 is made tobe semitransparent to scatter the light emitted from a microscope tooptically eliminate the outlines of the pores 4 of a filter 3. That is,optically speaking, the pores 4 are lit so that the degree of lightnessof the outlines of the pores 4 becomes similar to that of thesurrounding area, so that the pores 4 cannot be recognized. According tothe present invention, semitransparency is obtained by activelyrefracting and scattering the light (that is to say that the light beamsare prevented from propagating in the straight direction and they arebent in many directions), which is emitted from a microscipe while thetransmittancy of the light is still kept high. It is not obtained byreducing the transmittency of the light.

There may by many ways to do so. For example, it is accomplished byincluding in a slide a large number of parcticles which have suchspecific charateristics that the light tramsmittancy thereof is lowerthan that of a slide or the light refraction thereof is much larger thanthat of a slide. Of course, the particles could possesses bothcharacteristics at the same time. It is possible to adjust the degree ofsemitransparency by changing the roughness of the particles.

Another way to effectively refract and scatter the light of a microscopeis to position a semitransparent layer 2, on the surface of slide 1,such that the light refraction thereof is larger than that of the slide1.

The semitransparent layer 2 can be made by including in a transparentmacromolecule resin and a innumerable number of articles 6. Saidarticles being an inorganic material made of, for example, ceramics.

To further effectively refract the light of a microscope, the bordersurface of the slide 1 and the semitransparent layer 2 can be maderougher. Of course, the light refraction of the slide 1 and thesemitransparent layer 2 in this case must be made dissimilar. There aremany ways to make the border surface rougher. In one of them, forexample, the surface of a roller, that presses and produces a slide 1 ora semitransparent layer 2, can be made rough.

A specimen preparation according to the present invention can beprepared by placing a filter 3 with material thereon on asemitransparent slide. To make a more complete preparation, an organicsolvent should be applied on the semitransparent slide, then the filter3 with the material 5 thereon, a mounting medium, and a cover glass 8should respectively be placed thereon. This method prevents theformation of air bubbles in the specimen preparation.

According to the present invention, when material on a filter 3 which isplaced on a semitransparent slide is examined with a microscope, theslide is illuminated from underneath by the light of a microscope. Insuch circumstances, since the slide 1 is arranged to be semitransparent,the light emitted thereto is refracted and scattered in variousdirections. The refracted light emits pores 4 of the filter 3 fromvarious directions, so that the degree of the lightness of the pores 4becomes similar to that of the surrounding area, resulting in theoutlines of the pores 4 being optically eliminated, though theyphysically still exist.

In the prior art, a slide is transparent and the light transmittancythereof is high, therefore the light of a microscope advances straightahead so that the pores 4 of a filter 3 are clearly observed, making theaccurate examination of material 5 impossible. According to the presentinvention, such a problem is eliminated since the light from amicroscope refracts and scatters in many directions.

As stated previously, the effective light refraction can be obtained bymaking the border surface of a slide 1 and a semitransparent layer 2 arough surface 7, because in this way, the light first refracts at therough surface 7 and again at the semitransparent layer 2. According tothis method, since the light can be refracted effectively, thesemitransparent layer 2 can be made thinner and the number of particlesincluded in the semitransparent layer 2 can be minimized, which isadvantageous when productivities are concerned.

A semitransparent slide according to the present invention can be mademechanically stronger and therefore it is used for a longer time. Thiscan be accomplished by making the semitransparent layer 2 with amacromolecule resin and the particles 6 included in a semitransparentlayer 2 with inorganic materials such as ceramics. By doing so, thechemical, heat and weather resistance of the resin of thesemitransparent layer 2 improves. As it is known, the resin can easilybe faded by chemicals and repeated use. Also, the mechanical strength ofthe resin is weaker than glass. Therefore, to include inorganicparticles 6 in the resin to eliminate the weakness of the resin isextremely effective to make the semitransparent slide as strong as aslide made of glass.

BEST MODE FOR CARRYING OUT OF THE INVENTION

The details of the present invention are described hereinafter. FIG. 1shows a preferred embodiment of the present invention. In the preferredembodiment, an innumerable number of particles, made of ceramics,globular shaped powder, pigments, powdered bones and so on, having alight refraction dissimilar to the slide 1 and a characteristic of aremarkable light refraction, are included in the material of slide 1 tomake the transparent slide 1 a muddy white color. The light emitted froma microscope is refracted inside the semitransparent slide, and therefracted light emits the pores 4 of a filter 3 from various directions,so that the pores 4 cannot be recognized when material on the filter 3are examined. The slide 1 can be colored other than white such as lightblue.

FIG. 2 shows another preferred embodiment of the present invention. Asemitransparent layer 2, in this case, is positioned on a transparentslide 1 made of macromolecule resin. The method of making themacromolecule resin semitransparent is not limited but, for example, maybe done by crystallizing the macromolecule to white or by blending otherresins in the macromolecule resin.

According to this preferred embodiment, the light is refracted by thesemitransparent layer 2 and the similar effect pursued by the firstpreferred emobodiment can be achieved. This semitransparent layer 2 iscrystallized in white and an innumerable number of particles 6 ofceramics are contained therein, so that the refraction and thescattering of the light can effectively be accomplished. By adding anemitting material or a layer to the surface of each particle, the lightcan more effectively be refracted and scattered.

FIG. 3 shows an another preferred embodiment of the present invention,wherein the border surface of the slide 1 and the semitransparent layer2 is made to be a rough surface 7. Thus, the light is refracted by therough surface 7, and it is further refracted by the semitransparentlayer 2, so that the light can actively be refracted and scattered.Also, the slide 1 and the semitransparent layer 2 can more firmly stickto each other.

The rough surface 7 can be made by blasting sand on the surface of theslide 1 or by soaking the slide 1 in chemicals such as a fluoride acid.

The semitransparent layer 2 can be positioned on the surface of theslide 1 by such methods as wherein the macromolecule resin is pressedand hardened on the slide 1, applied or sprayed to the surface of theslide 1, transcribed onto the slide 1, or the slide 1 is soaked in themacromolecule resin. In this case, for example, the thickness of theslide 1 may be about 1 mm and that of the semitranparent layer 2 lessthan 0.2 mm to obtain a total thickness of the semitransparent slideless than 1.2 mm to meet the requirements of JIS (Japanese IndustrialStandard).

To obtain the object of the present invention, a high transmittancy andan active refraction and acattering are required by a transparent slide.The semitransparent slide described above possesses more than 60˜80percent of transmittancy and a highly active light refraction caused bythe semitransparent layer 2, so that the pores 4 of the filter 3 can becompletely erased optically. It is a highly remarkable slide.

The shape of the semitransparent slide according to the presentinvention is not limited to those described as the preferredembodiments. A semitransparent slide can be arranged such as asandwich-like shape by making another transparent layer on thesemitransparent layer 2. In this case, the thickness of thesemitransparenet layer 2 may be prepared to be about 0.2 mm and that oftransparent layer may be about 0.01 mm, making the total thickness ofthe semitransparent slide, including the transparent slide 1 about 1.2mm. By making the semitransparent slide a sandwich-like shape andadjusting the thickness of the transparent layer, the resolution becomesbetter and the outlines of material can clearly be recognized, so thatan examination of the material becomes easier and accurate.

Further, a semitransparent layer 2 can be positioned not only on the topsurface of the slide 1 but also on the under surface, or both top andunder surfaces of the slide 1. The characteristics of the presentinvention are that a slide 1 is made to be semitransparent toefficiently refract and scatter the light emitted from a microscope tooptically erase the pores 4 of a filter 3. The method of making theslide semitransparent is not limited at all.

Semitransparent as described herein includes one that looks opaque bynaked eyes but transmits light therethrough.

A slide 1 here includes such that can place things such as material andfilter 3 thereon, and it is not limited to a glass slide. Therefore, itincludes, for example, products of macromolecule resin and ceramics.

Further, a slide here includes one that can be recognized as opaque ortransparent depending on the visual angle.

FIG. 4 shows a specimen preparation made with a semitransparent slideaccording to the present invention and a filter 3. It is prepared by thefollowing procedures. First, xylol is applied on the semitransparentslide and a filter 3 with material thereon is placed on it. The xylolflows into the pores 4 and pushes the air out of the pores 4. Therefore,the whole material 5 can be positioned at the same level on the filter 3and air bubbles cannot be formed, so that examination of the materialbecomes easy and accurate.

Then, a mounting medium 9 is applied to the filter 3 and a cover glass 8is placed thereon, completing the procedure. At this point, the xylolapplied previously evaporates and it is replaced by the mounting medium9, so that the filter 3 and the slide 1 are firmly stuck to each other,and the material 5 can be completely sealed.

The method of making a preparation is not limited to the one explainedabove. For example, any other appropriate mediums such as immersion oilcan be accordingly applied thereon.

A characteristic of a method of making a preparation according to thepresent invention is that where a filter 3 with material collectedthereon is placed on the semitransparent slide. The effect is that thelight emitted from a microscope is actively refracted and scattered, sothat the pores 4 of the filter 3 are optically erased, making theaccurate and easy examination of the material 5 possible.

The material of a filter 3 is not limited. A polyester membranfilter, apolycarbonate membrane filter, having a thickness thereof of about 10 μmwith an innumerable number of pores 4 therethrough, the diameter of thepores 4 being a few micron, and so-called a honeycomb-like filter whichis made of a thin alumina having innumerable number of honeycomb-likepores therethrough are some of the filters preferably used as a membranefilter 3.

FIG. 5 shows a cover glass 8 according to the present invention. A metalevaporation layer 10, the thickness thereof being in microns toangstroms and is formed of metal molecules such as magnesium fluoride,aluminum and zirconium oxide, is placed on the transparent cover glass 8by a vacuum evaporation, so-called multicoating. According to a test,this cover glass is especially effective when material is to be examinedat high magnification, since the multi coating layer, the metalevaporation layer 10, prevents the light reflection when the light comesout from the cover glass 8 to the air. This improves the lighttransmittancy of the cover glass 8.

As stated previously, according to the present invention, a slide ismade to be semitransparent, so that the outlines of the pores of afilter cannot be observed. Therefore, neither a so-called transcriptionor a melting procedure become unnecessary, and material on the filtercan be recognized clearly.

By arranging the border surface between the slide and thesemitransparent layer, the light emitted from a microscope can be moreeffectively refracted and scattered.

By making a semitransparent layer with a macromolecule resin andincluding inorganic particles therein, the degree of the lightrefraction becomes higher, the chemical, heat and whether resistanceimproves, and mechanical strength advances, so that the durability ofthe semitransparent slide becomes as tough as the one made of glass.

According to the method of making a specimen preparation, a filter withmaterial thereon is placed on a semitransparent slide, which eliminatesthe need for a smearing process, so that the material cannot bedestroyed or deformed and they do not even drop in a dyeing from thefilter. This enables you to examine the material without error.

Another effect of the method of making a preparation according to thepresent invention is that it does not require either a lot of expreienceor good skill and anyone could make the preparation without muchdifficulty. This improves the efficiency of preparing specimenpreparations.

Further, by making a metal evaporation particle layer on a cover glass,material can be examined more clearly especially when they arerecognized at high magnification.

I claim:
 1. A slide and filter combination for a microscope comprising:asemitransparent slide comprising one or a plurality of layers such thatsaid slide has two opposed outer flat surfaces; and a filter having aporous surface placed adjacent to one of said opposed outer flatsurfaces of said semitransparent slide such that light of a microscopeis transmitted through said slide and said filter to permit observanceof a specimen placed on said filter by a user; wherein at least onelayer of said slide has a semitransparent portion including means forrefracting and scattering light transmitted therethrough such that poresof the filter are not visible to a user.
 2. The combination according toclaim 1, wherein said slide comprises a layer which is crystallized toappear white.
 3. The combination according to claim 1, wherein saidsemitransparent portion is composed of a macromolecule beingcrystallized to appear milky in color.
 4. The combination according toclaim 1, wherein the semitransparent slide comprises a transparent layercomposed of a transparent material and a semitransparent layer opposedadjacent to said transparent layer, said semitransparent layer composedof a transparent material containing particles embedded therein, saidparticles having a lower light transmittance than said transparentmaterial, said particles refracting and scattering light transmittedthrough said semitransparent layer.